摘要：
Adventitious root (AR) formation is a critical process for plant clonal propagation. The role of plant secondary metabolites in AR formation is still poorly understood. Chemical and physical mutagenesis in combination with somatic variation were performed on Artemisia annua in order to obtain a mutant with changes in adventitious rooting and composition of plant secondary metabolites. Metabolic and morphological analyses of the iar (increased adventitious rooting) mutant coupled with in vitro assays were used to elucidate the relationship between plant secondary metabolites and AR formation. The only detected differences between the iar mutant and wild-type were rooting capacity and borneol/camphor content. Consistent with this, treatment with borneol in vitro promoted adventitious rooting in wild-type. The enhanced rooting did not continue upon removal of borneol. The iar mutant displayed no significant differences in AR formation upon treatment with camphor. Together, our results suggest that borneol promotes adventitious rooting whereas camphor has no effect on AR formation.
通讯机构：
[Lin, Yong] H;Hunan Agr Univ, Natl Res Ctr Engn & Technol Utilizat Bot Funct In, Changsha 410128, Hunan, Peoples R China.
关键词：
Mass spectrometry;Membrane proteomics;Sample preparation;Shotgun analysis;Sodium deoxycholate
摘要：
In current shotgun-proteomics-based biological discovery, the identification of membrane proteins is a challenge. This is especially true for integral membrane proteins due to their highly hydrophobic nature and low abundance. Thus, much effort has been directed at sample preparation strategies such as use of detergents, chaotropes, and organic solvents. We previously described a sample preparation method for shotgun membrane proteomics, the sodium deoxycholate assisted method, which cleverly circumvents many of the challenges associated with traditional sample preparation methods. However, the method is associated with significant sample loss due to the slightly weaker extraction/solubilization ability of sodium deoxycholate when it is used at relatively low concentrations such as 1%. Hence, we present an enhanced sodium deoxycholate sample preparation strategy that first uses a high concentration of sodium deoxycholate (5%) to lyse membranes and extract/solubilize hydrophobic membrane proteins, and then dilutes the detergent to 1% for a more efficient digestion. We then applied the improved method to shotgun analysis of proteins from rat liver membrane enriched fraction. Compared with other representative sample preparation strategies including our previous sodium deoxycholate assisted method, the enhanced sodium deoxycholate method exhibited superior sensitivity, coverage, and reliability for the identification of membrane proteins particularly those with high hydrophobicity and/or multiple transmembrane domains.
作者：
Chen, H. X.;Jiang, H.;Huo, J. L.;Wen, J. F.*;Zhu, H. S.;...
期刊：
Genetics and Molecular Research,2014年13(2):2531-2538 ISSN：1676-5680
通讯作者：
Wen, J. F.
作者机构：
[Jiang, H.; Chen, H. X.] Hunan Agr Univ, Coll Hort, Changsha, Hunan, Peoples R China.;[Lv, J. H.; Ma, C. H.; Zhu, H. S.; Huo, J. L.; Deng, M. H.; Zhou, H.] Yunnan Agr Univ, Kunming, Peoples R China.;[Wen, J. F.] Kunming Univ Sci & Technol, Kunming, Peoples R China.
通讯机构：
[Wen, J. F.] K;Kunming Univ Sci & Technol, Kunming, Peoples R China.
摘要：
RATIONALE Alkaloids with significant therapeutic effects are the main active constituents of Macleaya cordata, which is a perennial herb plant in the Papaveraceae family. A systematic and novel method for speculating and identifying the structures of alkaloids in M. cordata fruits by high-performance liquid chromatography/quadrupole-time-of-flight mass spectrometry (HPLC/Q-TOF-MS) with a screening procedure was reported. METHODS Investigation of mass spectral fragmentation of alkaloids was carried out based on the tandem mass spectrometry (MS/MS) data analyses of eight reference substances. The skeletons of alkaloids were determined by their ultraviolet spectra (UV) and MS/MS data. The substituent groups of the alkaloids were acquired through a screening procedure developed in our laboratory and MS/MS data. The substituent linkage sites were deduced by MS/MS fragmentation behavior, as well as biosynthetic pathways of related alkaloids. RESULTS The structures of 21 alkaloids were speculated in this study, 10 of which were reported for the first time in M. cordata. Furthermore, benzyltetrahydroisoquinoline and N-methyltetrahydroprotoberberine-type alkaloids were discovered, which indirectly proved that the biosynthetic pathways of benzophenanthridine alkaloids reported in Eschscholtzia california existed in M. cordata as well. CONCLUSIONS HPLC/Q-TOF-MS combined with a screening procedure was a systematic and reliable method for speculating and elucidating the structures of alkaloids. This study might be useful for the identification of other compounds in herbal medicines. Copyright (c) 2014 John Wiley & Sons, Ltd.
摘要：
A new intact resin glycoside (3) and two glycosidic acids (1 and 2), all having a common trisaccharide moiety and (11S)-hydroxytetradecanoic acid or (3S,11S)-dihydroxytetradecanoic acid as the aglycone, were obtained from the roots of Porana duclouxii. Their structures were elucidated by spectroscopic analyses and chemical correlations. These compounds represent the first examples of resin glycosides from the genus Porana.
摘要：
The Fusarium oxysporum species complex consists of fungal pathogens that cause serial vascular wilt disease on more than 100 cultivated species throughout the world. Gene function analysis is rapidly becoming more and more important as the whole-genome sequences of various F. oxysporum strains are being completed. Gene-disruption techniques are a common molecular tool for studying gene function, yet are often a limiting step in gene function identification. In this study we have developed a F. oxysporum high-efficiency gene-disruption strategy based on split-marker homologous recombination cassettes with dual selection and electroporation transformation. The method was efficiently used to delete three RNA-dependent RNA polymerase (RdRP) genes. The gene-disruption cassettes of three genes can be constructed simultaneously within a short time using this technique. The optimal condition for electroporation is 10 mu F capacitance, 300 Omega resistance, 4kV/cm field strength, with 1 mu g of DNA (gene-disruption cassettes). Under these optimal conditions, we were able to obtain 95 transformants per mu g DNA. And after positive-negative selection, the transformants were efficiently screened by PCR, screening efficiency averaged 85%: 90% (RdRP(1)), 85% (RdRP(2)) and 77% (RdRP(3)). This gene-disruption strategy should pave the way for high throughout genetic analysis in F. oxysporum. (C) 2014 Elsevier GmbH. All rights reserved.
作者机构：
[邓近平; 杨玲媛] College of Animal Science and Technology, Hunan Agricultural University, Changsha 410128, China;[汤少勋; 周传社; 王敏; 谭支良] Institute of Subtropical Agriculture, Chinese Academy of Sciences, Changsha 410125, China;College of Horticulture Landscape, Hunan Agricultural University, Changsha 410128, China;[王荣] College of Animal Science and Technology, Hunan Agricultural University, Changsha 410128, China, Institute of Subtropical Agriculture, Chinese Academy of Sciences, Changsha 410125, China;[颜志成] Institute of Subtropical Agriculture, Chinese Academy of Sciences, Changsha 410125, China, College of Horticulture Landscape, Hunan Agricultural University, Changsha 410128, China
通讯机构：
[Deng, J.-P.] C;College of Animal Science and Technology, Hunan Agricultural University, Changsha 410128, China
摘要：
Cucurbitacins are triterpenoids that confer a bitter taste in cucurbits such as cucumber, melon, watermelon, squash, and pumpkin. These compounds discourage most pests on the plant and have also been shown to have antitumor properties. With genomics and biochemistry, we identified nine cucumber genes in the pathway for biosynthesis of cucurbitacin C and elucidated four catalytic steps. We discovered transcription factors Bl (Bitter leaf) and Bt (Bitter fruit) that regulate this pathway in leaves and fruits, respectively. Traces in genomic signatures indicated that selection imposed on Bt during domestication led to derivation of nonbitter cucurbits from their bitter ancestors.
作者：
Chalhoub, Boulos*;Denoeud, France;Liu, Shengyi;Parkin, Isobel A. P.;Tang, Haibao;...
期刊：
Science,2014年345(6199):950-953 ISSN：0036-8075
通讯作者：
Chalhoub, Boulos
作者机构：
[Vinh Ha Dinh Thi; Mestiri, Imen; Chalabi, Smahane; Jabbari, Kamel; Just, Jeremy; Lu, Yunhai; Arnaud, Dominique; Canaguier, Aurelie; Le Clainche, Isabelle; Chalhoub, Boulos; Chelaifa, Houda; Belcram, Harry] Univ Evry Val dEssone, Inst Natl Rech Agron INRA, Unite Rech Genom Vegetale, UMR1165, F-91057 Evry, France.;[Da Silva, Corinne; Denoeud, France; Bento, Pascal; Wincker, Patrick; Labadie, Karine; Alberti, Adriana; Correa, Margot; Noel, Benjamin; Bernard, Maria; Aury, Jean-Marc; Battail, Christophe] Commissariat Energie Atom CEA, Inst Genom IG, F-91057 Evry, France.;[Denoeud, France; Wincker, Patrick] Univ Evry Val dEssone, UMR 8030, F-91057 Evry, France.;[Denoeud, France; Wincker, Patrick] Ctr Natl Rech Sci CNRS, UMR 8030, Evry, France.;[Hu, Qiong; Wang, Xinfa; Tong, Chaobo; Liu, Shengyi; Hua, Wei] Chinese Acad Agr Sci, Key Lab Biol & Genet Improvement Oil Crops, Minist Agr Peoples Republ China, Oil Crops Res Inst, Wuhan 430062, Peoples R China.
通讯机构：
[Chalhoub, Boulos] U;Univ Evry Val dEssone, Inst Natl Rech Agron INRA, Unite Rech Genom Vegetale, UMR1165, 2 Rue Gaston Cremieux, F-91057 Evry, France.
摘要：
Oilseed rape (Brassica napus L.) was formed ∼7500 years ago by hybridization between B. rapa and B. oleracea, followed by chromosome doubling, a process known as allopolyploidy. Together with more ancient polyploidizations, this conferred an aggregate 72x genome multiplication since the origin of angiosperms and high gene content.We examined the B. napus genome and the consequences of its recent duplication. The constituent An and Cn subgenomes are engaged in subtle structural, functional, and epigenetic cross-talk, with abundant homeologous exchanges. Incipient gene loss and expression divergence have begun. Selection in B. napus oilseed types has accelerated the loss of glucosinolate genes, while preserving expansion of oil biosynthesis genes. These processes provide insights into allopolyploid evolution and its relationship with crop domestication and improvement.
关键词：
Rickettsia;Nymphs;Fluorescent in situ hybridization;Population density;Cotton;Population dynamics;Insects;Tomatoes
摘要：
Bacterial symbionts often enhance the physiological capabilities of their arthropod hosts and enable their hosts to expand into formerly unavailable niches, thus leading to biological diversification. Many arthropods, including the worldwide invasive whitefly Bemisia tabaci, have individuals simultaneously infected with symbionts of multiple genera that occur in different locations in the host. This study examined the population dynamics of symbionts that are located in different areas within B. tabaci. While densities of Portiera and Hamiltonella (which are located in bacteriocytes) appeared to be well-regulated during host development, densities of Rickettsia (which are not located in bacteriocytes) were highly variable among individual hosts during host development. Host mating did not significantly affect symbiont densities. Infection by Tomato yellow leaf curl virus did not affect Portiera and Hamiltonella densities in either sex, but increased Rickettsia densities in females. High and low temperatures did not affect Portiera and Hamiltonella densities, but low temperature (15 degrees C) significantly suppressed Rickettsia densities whereas high temperature (35 degrees C) had little effect on Rickettsia densities. The results are consistent with the view that the population dynamics of bacterial symbionts in B. tabaci are regulated by symbiont location within the host and that the regulation reflects adaptation between the bacteria and insect.
摘要：
A series of isoquinoline alkaloids including tetrahydroprotoberberines, N-methyl tetrahydroprotoberberines and protoberberines were facile synthesised with protopines as the starting material. All compounds were evaluated for their antibacterial activities against four pathogenic bacteria Escherichia coli, Staphyloccocus aureus, Staphylococcus gallinarum and Salmonella choleraesuis. Experimental results indicated that protoberberines were the most active compounds to the target bacteria among the tested alkaloids. It was suggested that planar molecule with high aromatisation level (e.g. coptisine 5 and berberine 6) or a positive charge of the molecules (e.g. N-methyl tetrahydroprotoberberines 11 and 12) had a positive influence on the antibacterial effects.
期刊：
JOURNAL OF ORGANIC CHEMISTRY,2014年79(22):11264-11269 ISSN：0022-3263
通讯作者：
Liang, Yun
作者机构：
[Huang, Hui; Wu, Lijun; Liang, Yun] Hunan Normal Univ, Natl & Local Joint Engn Lab New Petrochem Mat & F, Key Lab Chem Biol & Tradit Chinese Med Res, Minist Educ, Changsha 410081, Hunan, Peoples R China.;[Cheng, Pi] Hunan Agr Univ, Natl Res Ctr Engn Technol Utilizat Funct Ingredie, Changsha 410128, Hunan, Peoples R China.
通讯机构：
[Liang, Yun] H;Hunan Normal Univ, Natl & Local Joint Engn Lab New Petrochem Mat & F, Key Lab Chem Biol & Tradit Chinese Med Res, Minist Educ, Changsha 410081, Hunan, Peoples R China.
摘要：
A metal-free decarboxylative amination of 4-(ethoxycarbonyl)-2,3-allenols by TsNCO via base-induced aza-Michael addition/elimination has been developed. A variety of substituted N-tosyl 1,3-dien-2-yl amines were obtained in good yields and excellent regio- and stereoselectivity. Moreover, this transformation could be applied in preparation of 2-amino-trienes.
摘要：
BACKGROUND: Accurate evaluation of gene expression requires normalization relative to the expression of reliable reference genes. Expression levels of "classical" reference genes can differ, however, across experimental conditions. Although quantitative real-time PCR (qRT-PCR) has been used extensively to decipher gene function in the sweetpotato whitefly Bemisia tabaci, a world-wide pest in many agricultural systems, the stability of its reference genes has rarely been validated. RESULTS: In this study, 15 candidate reference genes from B. tabaci were evaluated using two Excel-based algorithms geNorm and Normfinder under a diverse set of biotic and abiotic conditions. At least two reference genes were selected to normalize gene expressions in B. tabaci under experimental conditions. Specifically, for biotic conditions including host plant, acquisition of a plant virus, developmental stage, tissue (body region of the adult), and whitefly biotype, ribosomal protein L29 was the most stable reference gene. In contrast, the expression of elongation factor 1 alpha, peptidylprolyl isomerase A, NADH dehydrogenase, succinate dehydrogenase complex subunit A and heat shock protein 40 were consistently stable across various abiotic conditions including photoperiod, temperature, and insecticide susceptibility. CONCLUSION: Our finding is the first step toward establishing a standardized quantitative real-time PCR procedure following the MIQE (Minimum Information for publication of Quantitative real time PCR Experiments) guideline in an agriculturally important insect pest, and provides a solid foundation for future RNA interference based functional study in B. tabaci.
